This study screened for radio-resistant strains lactic acid bacteria (LAB) by evaluating their capability to survive exposure to ionizing radiation. Ten strains of LAB - Lactobacillus bulgaricus, Lactobacillus paracasei, Lactobacillus casei, Lactobacillus acidophilus, Lactobacillus plantarum, Lactobacillus delbruekii, Lactococcus lactis, Streptococcus thermophilus, Bifidobacterium breve, and Pediocuccos pentosaceus - were selected and subcultuted twice. The LAB was then further cultured for 3 d at $37^{\circ}C$ to reach 7-10 Log colony-forming units (CFU)/mL prior to irradiation and immediately exposed to gamma rays or electron beams with absorbed doses of 0, 1, 2, 3, 4, 5, 6, 8, and 10 kGy. Gamma irradiation gradually decreased the number of the tested viable LAB, and the effect was irradiation dose dependent. A similar effect was found in electron beam-irradiated LAB. Radiation sensitivity of LAB was calculated as $D_{10}$ values, which ranged from 0.26 kGy to 0.9 kGy and 0.5 kGy to 1.44 kGy with exposure to gamma and electron beam irradiation, respectively, in all tested LAB. L. acidophilus was the most resistant to gamma and electron beam irradiation, with $D_{10}$ values of 0.9 kGy and 1.44 kGy, respectively. These results suggest that L. acidophilus might be suitable for the preparation of probiotics as direct-fed microbes for astronauts in extreme space environments.
Lactic acid is an important product arising from the anaerobic fermentation by lactic acid bacteria (LAB). It is used in the pharmaceutical, cosmetic, chemical, and food industries as well as for biodegradable polymer and green solvent production. The poly lactic acid (PLA) is an important material for bio-plastic manufacturing process. For PLA production by new LAB, we screened LAB isolates from shellfish. A total of 28 LAB were isolated from various shellfishes. They were all Gram positive, oxidase and catalase negative. Based on API 50CHL kit, 7 strains among the 28 isolates were identified as Lactobacillus plantarum, 6 strains as Lactobacillus delbrueckii, 5 strains as Leuconostoc mesenteroides, 3 strains as Lactobacillus brevis, 2 strains as Lactococcus lactis, 1 strain as Lactobacillus salivarius, 1 strain as Lactobacillus paracasei, 1 strain as Lactobacillus pentosus, 1 strain as Lactobacillus fermentum and 1 strain as Pediococcus pentosaceu. Also, we examined the amount of total lactic acid produced by these new strains by HPLC analysis with Chiralpak MA column. One strain E-3 from Mytilus edulis was indentified as Lactobacillus plantarum and found to produce 20.0 g/L of D-form lactic acid from 20 g/L of dextrose. Further studies are underway to increase the D-lactic acid production by E-3.
The aim of this study was to manufacture functional high protein fermented beverage, using whey protein concentrate (WPC) and Lactobacillus plantarum DK211 isolated from kimchi, and to evaluate the physicochemical, functional, and sensory properties of the resulting product. The fermented whey beverage (FWB) was formulated with whey protein concentrate 80 (WPC 80), skim milk powder, and sucrose; and fermented with Lactobacillus plantarum DK211 as single, or mixed with Lactococcus lactis R704, a commercial starter culture. The pH, titratable acidity, and viable cell counts during fermentation and storage were evaluated. It was found that the mixed culture showed faster acid development than the single culture. The resulting FWB had high protein (9%) and low fat content (0.2%). Increased viscosity, and antioxidant and antimicrobial activity were observed after fermentation. A viable cell count of 109 CFU/mL in FWB was achieved within 10 h fermentation, and it remained throughout storage at 15℃ for 28 d. Sensory analysis was also conducted, and compared to that of a commercial protein drink. The sensory scores of FWB were similar to those of the commercial protein drink in most attributes, except sourness. The sourness was highly related with the high lactic acid content produced during fermentation. The results showed that WPC and vegetable origin lactic acid bacteria isolated from kimchi might be used for the development of a high protein fermented beverage, with improved functionality and organoleptic properties.
Recently, poly-γ-glutamic acid synthetase A (pgsA) has been applied to display exogenous proteins on the surface of Lactobacillus casei or Lactococcus lactis, which results in a surface-displayed component of bacteria. However, the ability of carrying genes encoded by plasmids and the expression efficiency of recombinant bacteria can be somewhat affected by the longer gene length of pgsA (1,143 bp); therefore, a truncated gene, pgsA, was generated based on the characteristics of pgsA by computational analysis. Using murine IL-10 as an exogenous gene, recombinant Lactobacillus plantarum was constructed and the capacity of the surface-displayed protein and functional differences between exogenous proteins expressed by these strains were evaluated. Surface expression of IL-10 on both recombinant bacteria with anchorins and the higher expression levels in L. plantarum-pgsA'-IL-10 were confirmed by western blot assay. Most importantly, up-regulation of IL-1β, IL-6, TNF-α, IFN-γ, and the nuclear transcription factor NF-κB p65 in RAW264.7 cells after stimulation with Poly(I:C) or LPS was exacerbated after co-culture with L. plantarum-pgsA. By contrast, IL-10 expressed by these recombinant strains could reduce these factors, and the expression of these factors was associated with recombinant strains that expressed anchorin (especially in L. plantarum-pgsA'-IL-10) and was significantly lower compared with the anchorin-free strains. These findings indicated that exogenous proteins could be successfully displayed on the surface of L. plantarum by pgsA or pgsA', and the expression of recombinant bacteria with pgsA' was superior compared with bacteria with pgsA.
This study was conducted to confirm the availability of lactic acid bacteria as probiotics haying inhibitory effects to cancer cell line. Five lactic acid bacteria showing anti-cancer activity were compared by acid tolerance, bile tolerance, antibiotics resistance, milk fermentation, stability, and cell adherence activity to colon epithelial cell. The results obtained are as follows : In acid tolerance, all strains did not have a resistance below pH 3.0 and 3.5 except Lactobacillus plantarum KCTC3099. In antibiotics resistance, Lactococcus lactis and L. plantarum KCTC3099 were resistant to cotrimoxazol (128 mg/1), and Bifidobacterium adolescentis Maeil-K8 and B. infantis Maeil-K9 were resistant to doxycylin and gentamycin (4 mg/1). In case of cell adherence ability to Caco-2 cell, B. infantis Maeil-K9 was found to be superior to others as 3.1%, while the others were less than 0.5%. When the strains were cultured to milk base, viable counts of the strains tested increased more 1 log cycle than inoculation, but acid production was very low except L. plantarum KCTC3099. Also, L. plantarum KCTC3099, B. adolescentis Maeil-K8, and B. infantis Maeil-K9 were stable in fermented milk base during storage. In conclusion, L. plantarum KCTC3099 and B. infantis Maeil-K9 were confirmed to be superior for the availability as probiotics.
Seong, Eun-Hak;Lee, Myeong-Jong;Kim, Hojun;Shin, Na Rae
Journal of Korean Medicine for Obesity Research
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v.18
no.2
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pp.106-114
/
2018
Objectives: We investigated about the microbial properties and changes in the efficacy of the Codonopsis lanceolata (CL) by natural fermentation. Methods: CL was fermented for four weeks in a well-ventilated place with 2.5% salt. pH, total sugar, total polyphenol, and total flavonoid were measured to determine fermentation characteristics according to fermentation period and salt treatment. Polymerase chain reaction denaturing gradient gel electrophoresis and random amplification of polymorphic DNA-polymerase chain reaction were carried out for microbial analysis during fermentation. In addition, HepG2 cell was cultured to check the lipid accumulation through oil red O staining and the glucose uptake was analyzed by measuring the 2-NBDG at C2C12 cell. Results: The pH level and the total sugar decreased with the CL fermentation. Total polyphenol and flavonoid increased after CL fermentation. It was confirmed that Leuconostoc mesenteroides were maintained continuously during fermentation. In the salt treatment CL, there was a sharp increase in Rahnella aquatilis. Lactobacillus plantarum matrix was observed in fermented CL. In addition, Lactococcus lactis, Weissella koreensis, R. aquatilis, L. plantarum, Leu. mesenteroides have been added to the salt treatment. Glucose uptake were significantly increased after fermentation with salt for four weeks. Lipid accumulation in the HepG2 cells was observed that there was difference (P<0.01) between free fatty acid group (100%) and decreased 4 weeks after fermentation (90.38%) at $800{\mu}g/mL$. Conclusions: Total polyphenol and flavonoid were increased after CL fermentation. Especially, percentage of the glucose uptake and lipid accumulation inhibition increased in CL fermentation with salt. It is expected that fermentation of salt treated CL will be more effective in diabetes and fatty liver.
Development of expression vectors is important for the basic and applied researches on kimchi LAB (lactic acid bacteria). An expression vector, pSJE6c was constructed by inserting P6C promoter sequence from Lactococcus lactis into pSJE, a shuttle vector for E. coli and Leuconostoc species. To test the efficiency of pSJE6c, aga ($\alpha$-galactosidase) and lacZ ($\beta$-galactosidase) genes were expressed in Lactobacillus brevis 2.14. Compared to the pSJE, expression levels of both genes were increased, indicating P6C promoter was better than indigenous promoters. Enzyme activities of L. brevis cells harboring pSJE6caga (pSJE6c with aga) or pSJE6Z (pSJE6c with lacZ) were 1.5-2 fold higher than those with pSJEaga (pSJE with aga) or pSJEZ (pSJE with lacZ). More RNA transcripts were detected in cells harboring pSJE6c based recombinant plasmid. The results indicated that heterologous gene expressions in kimchi LAB could be improved significantly by use of efficient expression vectors.
This experiment was carried out to investigate the effect of dietary supplementation of complex probiotics and enzyme on air quality in finishing pig building and the performance of finishing pigs. A total 117 crossbred $[(Landrace{\times}Yorkshire){\times}Duroc]$ pigs were randomly arranged into nine groups and assigned to three treatments. Pigs were fed a basal diet supplemented with 0, 0.1% level of probiotics and 0.1% level of complex probiotics and enzyme until the market weight for 42 days of the experimental period. Ammonia and hydrogen sulfide concentrations in the finishing pig building were significantly (p<0.05) decreased by dietary supplementation of complex probiotics and enzyme compared with those of control, however, indoor carbon dioxide concentration was not affected by dietary supplementation of probiotics or complex probiotics and enzyme. Average daily feed intake and feed conversion ratio were significantly improved (p<0.05) with dietary supplementation of 0.1% complex probiotics and enzyme; however, average daily gain was not affected by dietary supplementation of probiotics or complex probiotics and enzyme. In conclusion, the results obtained from this experiment suggest that the dietary supplementation of complex probiotics and enzyme for finishing pigs may improve air quality in the finishing pig building and the performance.
Journal of the Korean Society of Food Science and Nutrition
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v.46
no.6
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pp.671-680
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2017
This study was conducted to examine the effects and potential mechanisms of action of black soybean extracts and fermented black soybean extracts by Lactobacillus rhamnosus GG (LGG) and Bifidobacterium animals subsp. lactis BB-12 (BB-12) on proliferation of human follicle dermal papilla cells (HFDPC). We examined changes in pH, total polyphenol, sugar, and reducing sugar contents according to fermentation period of black soybean extracts. Assay using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide was performed to determine cell toxicity levels of the four black soybean extracts [black soybean water extract (BWE), black soybean ethanol extract (BEE), fermented BWE (F-BEW), and fermented BEE (F-BEE)]. Changes in mRNA expression levels of hair growth promoting factors and hair growth inhibiting factors by the four black soybean extracts were measured by real-time PCR. In addition, phosphorylation levels of mitogen-activated protein kinase family proteins were measured by western blot analysis. As a result, fermentation of black soybeans significantly reduced pH, total polyphenols, and sugar/reducing sugar contents. All four black soybean extracts showed no cellular toxicity in HFDPC. In fact, BEE significantly enhanced cell viability of HFDPC at $100{\mu}g/mL$ compared to control. BWE, BEE, and BWE-F significantly increased mRNA expression of vascular endothelial growth factor, and all four extracts increased mRNA expression of fibroblast growth factor. However, mRNA expression levels of apoptosis-related genes were not affected by black soybean extracts in HFDPC. Furthermore, BWE, BEE, and BWE-F significantly increased phosphorylation levels of extracellular signal-regulated kinase compared to control. Taken together, we demonstrated that black soybean extracts enhanced proliferation of human follicle dermal papilla cells partially via activation of hair growth promoting factors, although no particular significant effects on proliferation were observed by fermentation of black soybeans.
The manufacturing process and quality properties of Lactococcus lactis-fermented yakchobugak (LFY) containing some colored agro-food products were investigated to develop a good organoleptic bugak from the leaf of the medicinal herb. The de-oiling rate of LFY by centrifugation was 37.6%, which was significantly different to the 3.5% value obtained after standing and the 8.9% value obtained with the beating method (p<0.05). Reducing power (RS) increased with addition of increasing levels of ethanol extracts from agro-food powders. The RS of green tea extract-supplemented LFY had the highest value of 0.97 at $500{\mu}g/mL$, and this was significantly different to values obtained using other extracts (p<0.05). At 83.84%, the DPPH (2,2-diphenyl-1-picrylhydrazyl) free-radical scavenging activity of green tea extract-supplemented LFY was the greatest of all samples tested, followed by LFY with woolgeom at 39.48%, LFY with black rice at 28.45%, and LFY with bokbunja at 22.24% all ethanol extracts were added at $50{\mu}g/mL$ (p<0.05). Acid values of green tea and bokbunja LFYs stored in transparent PE bags at $60^{\circ}C$ for 7 days were 1.82% and 2.03%, respectively. Textural hardness values of LFYs were less than 250.62, except for LFYs with black rice and woolgeom, but these values increased $2{\sim}3$-fold after lactic acid fermentation. Carbohydrate and protein content of LFYs were in the ranges $58.95{\sim}64.63%$ and $7.50{\sim}8.68%$, respectively. Lipid and calorie contents of woolgeom LFY had the lowest values of 22.02% and 490 Kcal, respectively.
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